Engineering Chinese hamster ovary cells to maximize sialic acid content of recombinant glycoproteins

• Published in: Nature biotechnology Vol. 17; no. 11; pp. 1116 - 1121
• Main Authors: Krummen, L, Weikert, S, Papac, D, Briggs, J, Cowfer, D, Tom, S, Gawlitzek, M, Lofgren, J, Mehta, S, Chisholm, V, Modi, N, Eppler, S, Carroll, K, Chamow, S, Peers, D, Berman, P
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.11.1999
• Subjects: Animals; Biological and medical sciences; Biotechnology; CHO Cells; Cricetinae; Fundamental and applied biological sciences. Psychology; Galactosyltransferases - metabolism; Genetic Engineering; Genetic technics; Glycoproteins - chemistry; Glycoproteins - genetics; Glycoproteins - metabolism; Methods. Procedures. Technologies; Modification of gene expression level; N-Acetylneuraminic Acid - chemistry; N-Acetylneuraminic Acid - metabolism; Rabbits; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Sialyltransferases - metabolism; Sialic acid; Enzyme; Transferases; Glycosyltransferases; Rodentia; Glycoprotein; Gene overexpression; N-Acetyllactosaminide α-2,3-sialyltransferase; Glycosylation; Recombinant cell; Gene expression; CHO cell line; β-N-Acetylglucosaminylglycopeptide β-1,4-galactosyltransferase; Vertebrata; Mammalia; Hexosyltransferases; Recombinant protein; Hamster
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/15104

We have engineered two Chinese hamster ovary cell lines secreting different recombinant glycoproteins to express high levels of human beta1,4-galactosyltransferase (GT, E.C. 2.4.1.38) and/or alpha2, 3-sialyltransferase (ST, E.C. 2.4.99.6). N-linked oligosaccharide structures synthesized by cells overexpressing the glycosyltransferases showed greater homogeneity compared with control cell lines. When GT was overexpressed, oligosaccharides terminating with GlcNAc were significantly reduced compared with controls, whereas overexpression of ST resulted in sialylation of >/=90% of available branches. As expected, GT overexpression resulted in reduction of oligosaccharides terminating with GlcNAc, whereas overexpression of ST resulted in sialylation of >/=90% of available branches. The more highly sialylated glycoproteins had a significantly longer mean residence time in a rabbit model of pharmacokinetics. These experiments demonstrate the feasibility of genetically engineering cell lines to produce therapeutics with desired glycosylation patterns.